AB 18:293-295 (2013)  -  DOI: https://doi.org/10.3354/ab00502

Improvement of single-cell gel electrophoresis (SCGE) alkaline comet assay

Xiang-Rong Xu1, Jun-Quan Zhu2, Ting Ye2, Chun-Lin Wang2, Yi-Feng Zhu2, Hans-Uwe Dahms3,**, Fan Jin1,**, Wan-Xi Yang4,*

1Department of Reproductive Endocrinology, Key Laboratory of Reproductive Genetics, Women?s Hospital, Zhejiang University School of Medicine, 1 Xueshi Road, Hangzhou 310006, China
2Key Laboratory of Applied Marine Biotechnology by the Ministry of Education, Ningbo University, Ningbo 315211, China
3Green Life Science Department, College of Natural Science, Sangmyung University, 7 Hongji-dong, Jongno-gu, Seoul 110-743, South Korea
4The Sperm Laboratory, College of Life Sciences, Zhejiang University, Hangzhou 310058, China
*Email: . **Co-corresponding author

ABSTRACT: Although the single-cell gel electrophoresis (SCGE) alkaline comet assay can detect DNA damage quickly and with high sensitivity, it does not work well for cryopreserved sperm of some marine teleosts. Using large yellow croaker Pseudosciaena crocea sperm that were cryopreserved in increasing concentrations (5 to 30%) of DMSO, we made modifications to the classic SCGE method that included using common microscopic slides without rough surfaces, pretreatment of sperm before gel spread, and a single layer of gel. Electrophoresis conditions were 130 mA, 15 V, and 60 min, providing a high SCGE sensitivity and definition of the comet image. The improved method is useful for detecting cryopreserved sperm DNA damage of some marine teleosts, especially for testing the quality of genetic resources.

KEY WORDS: Pseudosciaena crocea · Sperm · Cryopreservation · SCGE · DNA damage

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Cite this article as: Xu XR, Zhu JQ, Ye T, Wang CL and others (2013) Improvement of single-cell gel electrophoresis (SCGE) alkaline comet assay. Aquat Biol 18:293-295. https://doi.org/10.3354/ab00502

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